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. Author manuscript; available in PMC: 2013 Jan 1.
Published in final edited form as: Methods Mol Biol. 2012;821:349–359. doi: 10.1007/978-1-61779-430-8_22

Fig. 3.

Fig. 3

Knockdown of raptor, but not rictor or mTOR, activates Akt on Ser-473. We transfected U373:MG cells with control siRNA, siRNA against the mTORC2-associated mRNA rictor, the mTORC1-associated mRNA raptor, or to mTOR itself. Lysates were collected at 72 h and examined by immunoblot using antibodies shown. Knockdown of raptor blocked phosphorylation of rpS6 and increased the phosphorylation of Akt. In contrast, knockdown of rictor or mTOR both decreased the phosphorylation of Akt. Knockdown of rictor or raptor also induced autophagy, albeit to a lesser extent than that observed in response to knockdown of mTOR (as indicated by accumulation of LC3-II). (Reproduced from ref. 13 with permission from AAAS).